Wondered if anyone could recommend a good assay kit to measure cholesterol levels in cell extracts? I'm working with primary motor neurones so it needs to be quite sensitive as there will not be many cells to play with.

Thank you in advanceEmily

-EmilyG-

Speak to your nearest clinical laboratory. They'll have well established kits on automated platforms. They might object to running cell extracts. CSF would work but this may not be your sample matrix.

Other than that, its fairly easy to set up a colorimetric method. You'd need a cholesterase to hydrolyse the cholesterol esters. Then a cholesterol oxidase. The product of this is a cholesterone and peroxide. the peroxide is easily quantitated using aminoantipyrine and phenol in the presence of peroxidase. It makes an orange dye (quinoneimine) which you read at 510nm.